Biotypes B and Q of Bemisia tabaci and their relevance to neonicotinoid and pyriproxyfen resistance

Resistance monitoring for Bemisia tabaci field populations to the juvenile hormone mimic, pyriproxyfen, was conducted from 1996 to 2003 in commercial cotton fields in two areas of Israel: the Ayalon Valley (central Israel) and the Carmel Coast (northwestern Israel). Although the use of pyriproxyfen ceased in these areas in 1996-1997 (because of the resistance), resistance levels to pyriproxyfen declined to some extent in the fields but remained quite stable, and the susceptibility has not been totally restored. Two strains of B. tabaci collected from the Ayalon Valley in the late 1999 and 2002 cotton seasons (AV99L, AV02L) were assayed for their susceptibility to pyriproxyfen at F1, and subsequently a line of each strain was kept under controlled conditions without exposure to insecticides. After maintenance of more than 20 generations under laboratory conditions, the resistance to pyriproxyfen in the untreated strains substantially declined. This decline was concurrent with a replacement of Q biotype by B-type under non-insecticidal regimes; apparently B biotype was more competitive than the pyriproxyfen-resistant Q-type. Selection under controlled conditions with neonicotinoids on these B. tabaci strains resulted in continued pyriproxyfen resistance, predominantly of Q biotype. Based on our data, applications of either pyriproxyfen or neonicotinoids may select for biotype Q, which would survive to a greater degree where these insecticides are applied.     

西藏发现Q型烟粉虱

【目的】调查西藏自治区烟粉虱Bemisia tabaci(Gennadius)的发生情况。【方法】从西藏拉萨采集到烟粉虱各个虫态,采用3D数码显微镜观察所采集烟粉虱的形态特征,利用mt COⅠ分子标记检测烟粉虱的生物型。【结果】明确并详细描述了烟粉虱各形态特征,mt COⅠ分子标记检测显示西藏采集到的烟粉虱为Q生物型。【结论】在形态学鉴定的基础上,分子生物学鉴定该粉虱为Q型烟粉虱,这是Q型烟粉虱在西藏自治区发生的首次报道。     

Distribution of Bemisia tabaci (Hemiptera: Aleyrodidae) biotypes in North America after the Q invasion

After the 2004 discovery of the Bemisia tabaci (Gennadius) (Hemiptera Aleyrodidae) Q biotype in the United States, there was a vital need to determine the geographical and host distribution as well as its interaction with the resident B biotype because of its innate ability to rapidly develop high-level insecticide resistance that persists in the absence of exposure. As part of a coordinated country-wide effort, an extensive survey of B. tabaci biotypes was conducted in North America, with the cooperation of growers, industry, local, state, and federal agencies, to monitor the introduction and distribution of the Q biotype. The biotype status of submitted B. tabaci samples was determined either by polymerase chain reaction amplification and sequencing of a mitochondrial cytochrome oxidase I small subunit gene fragment and characterization of two biotype discriminating nuclear microsatellite markers or esterase zymogram analysis. Two hundred and eighty collections were sampled from the United States, Bermuda, Canada, and Mexico during January 2005 through December 2011. Host plants were split between ornamental plant and culinary herb (67%) and vegetable and field crop (33%) commodities. The New World biotype was detected on field-grown tomatoes (Solanum lycopersicum L.) in Mexico (two) and in commercial greenhouses in Texas (three) and represented 100% of these five collections. To our knowledge, the latter identification represents the first report of the New World biotype in the United States since its rapid displacement in the late 1980s after the introduction of biotype B. Seventy-one percent of all collections contained at least one biotype B individual, and 53% of all collections contained only biotype B whiteflies. Biotype Q was detected in 23 states in the United States, Canada (British Columbia and Ontario territories), Bermuda, and Mexico. Forty-five percent of all collections were found to contain biotype Q in samples from ornamentals, herbs and a single collection from tomato transplants located in protected commercial horticultural greenhouses, but there were no Q detections in outdoor agriculture (vegetable or field crops). Ten of the 15 collections (67%) from Canada and a single collection from Bermuda contained biotype Q, representing the first reports of biotype Q for both countries. Three distinct mitochondrial haplotypes of B. tabaci biotype Q whiteflies were detected in North America Our data are consistent with the inference of independent invasions from at least three different locations. Of the 4,641 individuals analyzed from 517 collections that include data from our previous work, only 16 individuals contained genetic or zymogram evidence of possible hybridization of the Q and B biotypes, and there was no evidence that rare hybrid B-Q marker co-occurrences persisted in any populations.     

不同作物田烟粉虱发生的时空动态

2008～2009年连续2年系统调查了番茄、茄子、棉花、大豆、玉米等寄主植物上烟粉虱种群发生的时间与空间动态。结果表明：不同寄主植物上的烟粉虱成虫及其伪蛹数量有显著性差异,其密度大小依次为：茄子>棉花>番茄>大豆>玉米。其中,在玉米上除了发现极少量的成虫逗留外,没有发现烟粉虱的卵及若虫。在发生的时间序列上,烟粉虱成虫及伪蛹的数量呈现为先逐渐上升后又下降的变化过程,发生高峰期集中在8月5日到8月31日,9月初以后烟粉虱数量慢慢减少。在空间分布上,表现为烟粉虱成虫喜食寄主的上部叶片。统计分析显示,寄主对烟粉虱成虫和伪蛹的数量的影响极显著,而年份对其数量的影响没有显著差异。由此得出的烟粉虱发生和达到高峰的时间,可为烟粉虱预测预报和区域性综合治理提供重要理论依据。     

Biotype-dependent secondary symbiont communities in sympatric populations of Bemisia tabaci

The sweet potato whitefly, Bemisia tabaci, harbors Portiera aleyrodidarum, an obligatory symbiotic bacterium, as well as several secondary symbionts including Rickettsia, Hamiltonella, Wolbachia, Arsenophonus, Cardinium and Fritschea, the function of which is unknown. Bemisia tabaci is a species complex composed of numerous biotypes, which may differ from each other both genetically and biologically. Only the B and Q biotypes have been reported from Israel. Secondary symbiont infection frequencies of Israeli laboratory and field populations of B. tabaci from various host plants were determined by PCR, in order to test for correlation between bacterial composition to biotype and host plant. Hamiltonella was detected only in populations of the B biotype, while Wolbachia and Arsenophonus were found only in the Q biotype (33% and 87% infection, respectively). Rickettsia was abundant in both biotypes. Cardinium and Fritschea were not found in any of the populations. No differences in secondary symbionts were found among host plants within the B biotype; but within the Q biotype, all whiteflies collected from sage harboured both Rickettsia and Arsenophonus, an infection frequency which was significantly higher than those found in association with all other host plants. The association found between whitefly biotypes and secondary symbionts suggests a possible contribution of these bacteria to host characteristics such as insecticide resistance, host range, virus transmission and speciation.     

不同生物型烟粉虱体内Wolbachia共生菌的检测及其系统树分析

Wolbachia是广泛分布于节肢动物体内一类共生细菌,它能够通过多种机制调控宿主的生殖方式。近年来的研究表明,Wolbachia与许多外来生物的成功入侵相关。本文利用长PCR的方法特异扩增了不同生物型烟粉虱（共24个种群）体内Wolbachia的wsp基因,结果发现B型和Q型烟粉虱入侵种群体内均未检测到Wolbachia,而在非B/Q型的浙江种群和肯尼亚种群体内检测到了Wolbachia。对该wsp基因进行测序并和已知序列进行同源性分析发现,浙江烟粉虱种群的Wolbachia属于B组Con/Rug亚种群,而肯尼亚种群属于B组Btab1亚种群。Wolbachia的存在与否可能与烟粉虱的成功入侵有一定的关系。图2表2参23     

辽宁省冬季温室内越冬粉虱伪蛹的种类鉴定及烟粉虱体内番茄黄化曲叶病毒的检测

为了揭示辽宁省冬季温室内越冬粉虱伪蛹的种类及烟粉虱Bemisia tabaci （Gennadius）携带番茄黄化曲叶病毒（tomato yellow leaf curl virus, TYLCV）情况, 于2012年1月份在辽宁省不同县市区的温室作物上采集了17份粉虱伪蛹样品（每样品含30头粉虱伪蛹） , 镜检鉴别粉虱种类并利用mtCOI基因对烟粉虱生物型进行了鉴定; 检测了烟粉虱携带TYLCV情况并对其PCR扩增产物进行了测序分析。结 果表明： 辽宁省冬季温室内存在越冬温室白粉虱Trialeurodes vaporariorum （Westwood）与烟粉虱。17份粉虱样品中, 11份样品为烟 粉虱样品, 6份样品为温室白粉虱和烟粉虱混合样品。混合样品中, 温室白粉虱仅在锦州凌海（LH）样品中占优势。17份烟粉虱样品（包 括混合样品）中, 仅有4份样品为B型与Q型混合样品, 其他13份样品烟粉虱生物型均为Q型。17份烟粉虱样品中有3份Q型烟粉虱样品检测 到TYLCV, 系统树分析进一步证实该病毒是TYLCV。调查结果为辽宁省粉虱与TYLCV的早期测报和防控提供了科学依据。     

New insights into the mitochondrial phylogeny of the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) in the Mediterranean Basin

The whitefly Bemisia tabaci is vector of plant infecting viruses and it is considered as one of the most important agricultural pests around the Mediterranean basin. At present, five biotypes of B. tabaci have been reported in the Mediterranean Basin: B, Q, S, T and M. To establish the phylogeographic relationship of these Mediterranean biotypes with others, 54 samples collected in Europe and Africa were analysed by sequencing the mitochondrial cytochrome oxidase I gene (mtCOI). The phylogeny showed that Spanish samples corresponding to the biotype S were related to the haplotype Uganda 2 of the African clade, associated with recent epidemic upsurges of cassava mosaic virus (CMD) in that country. This phylogeographic relationship gave support to a distinct subgroup revealed within the African clade. Bemisia tabaci collected from Euphorbia plants in Italy (biotype T) formed one of the three distinct subgroups existing within the Southeast/Far East Asian clade, while samples from Turkey (biotype M) clustered together with reference mitochondrial sequences from whiteflies from Pakistan and Thailand. Recent reports indicate that Bemisia populations corresponding to the biotypes S and T are distributed in areas larger than those initially delimited. Other results indicated that samples collected in Sudan grouped within the Mediterranean–North Africa clade together with reference sequences of the biotype Q corresponding to insects collected in Spain and Morocco. Mitochondrial haplotypes of B. tabaci samples collected on sweet potato in Ghana clustered with reference sequences of samples from Cameroon corresponding to one of the five Sub-Saharan subgroups already described in the African clade. These data extends the phylogenetic information of the B. tabaci species complex and present new questions to be investigated.     

Modeling evolution of resistance to pyriproxyfen by the sweetpotato whitefly (Homoptera: Aleyrodidae)

We used computer simulations to examine evolution of resistance to the insect growth regulator (IGR) pyriproxyfen by the sweetpotato whitefly, Bemisia tabaci (Gennadius), biotype B [=Bemisia argentifolii (Bellows & Perring)]. Consistent with trends seen in cotton (Gossipyium spp.) fields in Arizona and Israel, results suggest that evolution of resistance to pyriproxyfen may occur rapidly in this haplodiploid insect. Similar to results from models of diploid insects, resistance evolved faster with increases in toxin concentration, dominance of resistance in females, the initial frequency of the resistance allele, and the proportion of the region treated with pyriproxyfen. Resistance was delayed by fitness costs associated with resistance. Movement between treated and untreated cotton fields had little effect, probably because untreated cotton leaves provided internal refuges in treated fields and whiteflies were controlled with other insecticides in external refuges. Resistance evolved faster when susceptibility to pyriproxyfen was greater in susceptible males than susceptible females. In contrast, resistance evolved slower when susceptibility to pyriproxyfen was greater in resistant males than resistant females. Results suggest that growers may be able to prolong the usefulness of pyriproxyfen by applying lower toxin concentrations and promoting susceptible populations in refuges.     

云南Q型烟粉虱种群的鉴定

利用mtDNACOⅠ基因片段作标记,对采自云南昆明一品红上的烟粉虱Bemisiatabaci(Gennadius)种群生物型进行了鉴定。结果表明,该种群COⅠ基因片段(501bp)与摩洛哥Q型、西班牙Q型烟粉虱的相应碱基序列具有极高的同源性,与2种Q型烟粉虱的COⅠ基因仅分别相差3个和4个碱基,同源性分别达到994%和992%。序列分析表明我国云南昆明一品红上存在Q生物型烟粉虱     

中国部分地区烟粉虱生物型种类及系统发育关系分析

烟粉虱是一种危害严重的世界性害虫,是一个快速进化的复合种.本文利用mtCOI分子标记方法,对2010和2011年采自我国9个省(市)的33个烟粉虱种群进行了生物型鉴定和系统发育分析.结果表明： 我国目前存在着B型、Q型、ZHJ-1型、ZHJ-3型、An型以及Nauru型等6种生物型,且不同生物型的分布是不均匀的.遗传距离及系统发育树的分析结果显示,海南省的An型和台湾省的An型聚为一支,为同一来源;中国B型与来自法国和乌干达的B型的亲缘关系较近,同源性达到99%以上;中国的Q型与来自摩洛哥和法国的Q型聚为一个分支,而来自以色列和土耳其的Q型烟粉虱单独聚为一支,说明中国的Q型烟粉虱与来自地中海西部的Q型烟粉虱亲缘关系更近,可以推断中国的Q型烟粉虱的起源地为地中海西部地区.
     

烟粉虱抗药性的研究进展

烟粉虱是一种世界性的重要害虫,目前已对有机磷、拟除虫菊酯和氨基甲酸酯等许多常规杀虫剂产生高水平抗性,在一些地区烟粉虱对昆虫生长调节剂和烟碱类杀虫剂的抗性也是十分普遍.文中综述了世界范围内烟粉虱的抗药性现状、抗药性机理、以及抗药性与生物型的关系,并介绍了美国和以色列较为成功的烟粉虱抗药性治理计划.     

Utility of MtCOI polymerase chain reaction-restriction fragment length polymorphism in differentiating between Q and B whitefly Bemisia tabaci biotypes

The invasive, insecticide-resistant, Q whitefly biotype, has gradually spread to other countries including the US via human-mediated movement of plant materials. We assessed the utility of the VspI -based mtCOI (mitochondrion cytochrome oxidase I) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique as a rapid, cost-effective, and reliable alternative for differentiating the Q from the dominant B biotype in Arizona. Using the standard mtCOI gene sequencing and mtCOI PCR-RFLP techniques, we biotyped eight whitefly strains of five individuals each collected from poinsettia and cotton at different locations in Arizona. Complete concordance was observed between the two methods, with three strains being identified as the Q biotype and five samples as the B biotype. We also scanned the mtCOI gene sequences for VspI polymorphisms in the B and Q biotype whiteflies currently available in the GenBank database. This global screening revealed the existence of three and four VspI polymorphic types for the Q and B biotypes, respectively. Nevertheless, all three VspI polymorphic Q biotype whiteflies shared a common and unique VspI site that can be used to differentiate Q biotype from the four VspI polymorphic B biotype whiteflies identified. These results demonstrate that the VspI -based mtCOI gene PCR-RFLP provides a reliable diagnostic tool for differentiating the Q and B biotype whiteflies in the US and elsewhere.     

Use of mitochondrial cytochrome oxidase I polymerase chain reaction-restriction fragment length polymorphism for identifying subclades of Bemisia tabaci Mediterranean group

The Mediterranean group (commonly known as Q biotype; hereafter MED) of the sweetpotato whitefly, Bemisia tabaci (Gennadius), originated in the Mediterranean region, but it now has been found in at least 10 countries outside the Mediterranean. Collections of B. tabaci from some of these countries exhibit different pest behaviors and pesticide resistance characteristics, yet all may be classified as MED. A phylogenetic analysis of 120 mitochondrial cytochrome oxidase I (mtCOI) sequences (JN966761-JN966880) of MED whiteflies collected in Arizona and of 417 retrieved from the GenBank database resolves the MED into five subclades, designated as Q1-Q5. Only subclades Q1 and Q2 have been detected in the United States. Q1 and the other four subclades (Q2-Q5) differ in the number or position of the AluI recognition sites. Based on the differences in the AluI recognition sites reported here and the previously reported differences in VspI recognition sites, we developed a simple diagnostic technique to identify subclades Q1-Q5 by using mtCOI polymerase chain reaction (PCR)-restriction fragment-length polymorphism (RFLP). A test of a worldwide collection of whiteflies demonstrates that this combination mtCOIPCR-RFLP technique can reliably distinguish not only the MED from the Middle East-Asia Minor 1 group but also the Q1 from any of the other four MED subclades.     

Analysis of genetic diversity among different geographical populations and determination of biotypes of Bemisia tabaci in China

Abstract:  Analysis of the genetic diversity among 27 different geographical populations of Bemisia tabaci and determination of biotypes of B. tabaci in China based on amplified fragment-length polymorphism (AFLP) and the mitochondrial cytochrome oxidase I (mtDNA COI) gene sequences were conducted. In AFLP assay, the use of five primer combinations selected from 64 primer combinations allowed the identification of 229 polymorphic bands (97.03%) from 60 to 500 bp, suggesting abundant genetic diversity among different geographical populations of B. tabaci. To further identify biotypes of B. tabaci in China, the mtDNA COI gene sequences of nine representative populations from China, Israel and Spain were obtained. Molecular phylogenetic tree based on AFLP and mtDNA COI gene analyses revealed the presence, in China, of at least four different genetic groups of B. tabaci. B biotype, Q biotype and two non-B/Q biotype. B biotype was distributed nationwide. Q biotype was present only in the local region of China including the YunNan province and BeiJing city. This was also the first report about the invasion of Q biotype into China. Of the other two non-B/Q biotype groups, one was found in ShanDong and HeBei provinces, and another in ZheJiang province. The non-B/Q biotype ZheJiang population showed very high similarity with another Asian population India-IW ( AF110704 ) in mtDNA COI sequences and was possibly a Chinese indigenous population. The close monitoring of the Q biotype in locales of China where commercial plants were exported or imported, is now essential to avoid the further accidental distribution of the Q biotype.     

Species within the Bemisia tabaci (Hemiptera: Aleyrodidae) complex in soybean and bean crops in Argentina

The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a cryptic species complex that contains some of the most damaging pests in tropical and subtropical regions. Recent studies suggested that this complex is composed of at least 24 distinct species. We use the approach from these studies to consider the identity of B. tabaci in Argentina. Previous studies have suggested the presence of a B. tabaci presumably indigenous to the Americas and referred to as the BR biotype in Argentina. We placed the entity referred to as the BR biotype within the B. tabaci cryptic species complex using whiteflies collected in soybean and bean crops in northern and central Argentina. The whiteflies were assigned using the mitochondrial cytochrome oxidase (mtCOI) gene. Four unknown haplotypes plus two Argentina sequences from GenBank formed a cluster that was basal to the rest of the New World sequences. These sequences diverged from the consensus sequence across the range of 3.6 to 4.3%. Applying the species assignment rules of recent studies suggests that the individuals from Argentina form a separate species. A fifth unknown haplotype fell within the New World putative species and formed a distinct cluster with haplotypes from Panama. These results suggest that Argentina has two indigenous species belonging to the B. tabaci cryptic species complex. Rather than using mtCOI sequencing for all B. tabaci collected, a simple random amplified polymorphic DNA-polymerase chain reaction diagnostic was used and tested along with previously published primers designed to work specifically with the BR biotype from Brazil. These primers were either unable to distinguish between the two indigenous members of the complex in Argentina or indicated a difference when none was evident on the basis of mtCOI sequence comparison.     

DNA markers for identifying biotypes B and Q of Bemisia tabaci (Hemiptera: Aleyrodidae) and studying population dynamics

The two most widespread biotypes of Bemisia tabaci (Gennadius) in southern Europe and the Middle East are referred to as the B and Q-type, which are morphologically indistinguishable. In this study various DNA markers have been developed, applied and compared for studying genetic diversity and distribution of the two biotypes. For developing sequence characterized amplified regions (SCAR) and cleaved amplified polymorphic sequences (CAPS) techniques, single random amplified polymorphic DNA (RAPD) fragments of B and Q biotypes, respectively, were used. The CAPS were investigated on the basis of nuclear sodium channel and the mitochondrial cytochrome oxidase I genes (mtCOI) sequences. In general, complete agreement was found between the different markers used. Analysis of field samples collected in Israel for several years, using these markers, indicated that the percentage of the Q biotype tends to increase in field populations as time progresses. This may be attributed to the resistance of the Q biotype to neonicotinoids and pyriproxyfen and the susceptibility of the B biotype to these insecticides.     

Upregulation of temperature susceptibility in Bemisia tabaci upon acquisition of Tomato yellow leaf curl virus (TYLCV)

Acquisition of plant viruses has various effects on physiological mechanisms in vector insects. Bemisia tabaci is the only known vector of Tomato yellow leaf curl virus (TYLCV), which is a serious virus affecting tomato cultivars. In this study, the lifespan of Q1 biotype was compared between non-viruliferous (NV) and TYLCV-viruliferous (V) whiteflies. Total lifespan from egg to adult death of NV whiteflies was 62.54days but 10.64days shorter in V whiteflies. We investigated the temperature susceptibility of B. tabaci by comparing mortalities as well as heat shock protein (hsp) mRNA levels between NV and V whiteflies. For this, NV and V whiteflies were exposed for either 1 or 3h at 4, 25, and 35°C. The mortality of V whiteflies was higher than NV ones following exposure at either 4 or 35°C, but there was no significant difference at 25°C. Analysis of the expression level of heat shock protein (hsp) genes using quantitative real-time PCR showed that both cold and heat shock treatments stimulated higher expression of hsps (hsp40, hsp70, and hsp90) at various rates in V whiteflies than NV ones, but there was no difference at 25°C. All together, our results show that TYLCV acquisition accelerated the developmental rate and increased susceptibility to thermal stress in B. tabaci. Therefore, this modification may result in reduced vector longevity due to increased metabolic energy utilization. Our results provide insights into the complex interaction between vector fitness and thermal stress in relation to the acquisition and transmission of plant viruses.     

Silicon influence on resistance induction against Bemisia tabaci biotype B (Genn.) (Hemiptera: Aleyrodidae) and on vegetative development in two soybean cultivars

The potential of populations of Bemisia tabaci (Genn.) to become resistant to insecticides has stimulated research into alternative tactics of integrated pest management such as the induction of host-plant resistance. Recent data have shown that silicon can increase the degree of resistance of host plants to insect pests. Therefore the aim of our work was to study the effects of silicon application on the vegetative development of soybean plants and on the induction of resistance to the silverleaf whitefly, B. tabaci biotype B. We performed choice and no-choice tests of oviposition preference on two soybean cultivars, IAC-19 (moderately resistant to B. tabaci biotype B) and MONSOY-8001 (susceptible), with and without application of silicon. Silicon did not affect silverleaf whitefly oviposition preferences, but caused significant mortality in nymphs. Thus, silicon increased the degree of resistance to silverleaf whitefly. Silicon decreased the production of phenolic compounds, but did not affect lignin production. However, when applied to cultivar IAC-19, it increased the production of non-protein organic nitrogen. Silicon had no effect on the vegetative development of soybean plants, but it increased the degree of resistance to the silverleaf whitefly. We conclude that silicon applications combined with cultivar IAC-19 can significantly decrease silverleaf whitefly populations, having a positive impact both on the soybean plant and on the environment.     

Molecular characterization of Bemisia tabaci populations in Tunisia: genetic structure and evidence for multiple acquisition of secondary symbionts

A survey was conducted during 2009-2010 seasons to identify the distribution of Bemisia tabaci (Gennadius) biotypes in Tunisia. The genetic affiliation of collected populations was determined by polymerase chain reaction (PCR)-restriction fragment-length polymorphism (TaqI) of the mitochondrial cytochrom oxidase I (mtCOI) gene. Results, validated by sequencing and phylogenetic analysis, allowed the clustering of sampled sweetpotato whiteflies into B and Q biotypes. As B. tabaci harbors the obligatory bacterium Portiera aleyrodidarum, and a diverse array of secondary symbionts including Rickettsia, Hamiltonella, Wolbachia, Cardinium, Arsenophonus, and Fritschea, we report here the infectious status of Tunisian populations by secondary symbionts to find out a correlation between bacterial composition to biotype. The genetic variability and structure of B. tabaci populations in Tunisia was driven by analysis of molecular variance (AMOVA) and the hypothesis of isolation by distance was explored. Selective neutrality and genetic haplotype network tests suggested that Tunisian sweetpotato whiteflies have been undergoing a potential expansion followed by gene flow restriction.